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[Comparison of different methods in order to identify Proteus spp].

Comparison of different methods in order to identify Proteus spp. The objectives were: (a) to identify Proteus strains to species level, following Farmer's and O'Hara's conventional biochemical reactions; b) to evaluate the sensitivity and specificity of both the API 20E method and a schema of reduced reactions (TSI and MIO agar: motility, indole and ornithine) comparing them with conventional methodology, and c) to evaluate the utility of SDS-PAGE (total proteins) in order to identify Proteus strains to species level. Two hundred and five Proteus spp. clinical isolates, were collected between January 1998 and September 2004, from inpatients and outpatients at Hospital de Clinicas. Strains were identified by means of conventional methodology, the API 20E method, and a schema of reduced reactions. SDS-PAGE (total proteins) was used in 48 out of the 205 strains. The API 20E method identified 79 out of 87 (90.8%) strains of P. mirabilis, 103 out of 103 P. vulgaris complex, and 15 out of 15 P. penneri. Eight strains of P. mirabilis were identified as Proteus spp., the acid production from maltose being necessary to identify them to species level. The schema of reduced reactions identified 205 out of 205 (100%) strains, that is, this schema of reduced reactions identified all the strains to species level without any additional tests, in marked contrast to the API 20E method. The SDS-PAGE (total proteins) identified the three species of the genus, even if the strains of P. mirabilis showed different biochemical reactions.

Bacterial Typing Techniques↗

[Change in the quantity of coliform and Proteus bacteria during the ripening of raw, nonperishable meat products].

Studied were the changes in the amount of coliform and Proteus bacteria in the production of raw-dried and raw-smoked unperishable meat products, employing model testing procedures of experimentally contaminated meat products with Escherichia coli and Bacterium proteus, through following up the changes in the counts of coliform and Proteus bacteria in batches of regular production, and by the investigation of raw meat samples of varying moisture content. The studies were carried out at eight enterprises working under different technologic conditions. A total of twenty series of model experiments were carried out with artificially contaminated batches of products as well as twenty-one series of experiments with batches of regular production, and some 93 meat products, dried to a various extent, were sampled. Results showed that during the ripening process with unperishable meat products the reduce in the numbers of coliform bacteria is more intense than the decrease in the Proteus organisms. The process of smoking did not change the amount of these microbes. In unperishable raw meat products that meet the requirements of the Bulgarian State Standard coliforms are found in 43 per cent of the samples, and Proteus bacteria are not present.

Animals↗

[Histamine formation by Proteus species in tunafish (author's transl)].

Introducing the authors report on cases of scombroid poisoning episodes in food serving establishments. These cases were related to the formation of histamine in tuna meat due to Proteus morganii although the meat was sound when taken from cans. It is demonstrated experimentally (Table 1) and shown in a critical literature review (Table 2) that among Proteus species Proteus morganii is the only species being able of producing histamine levels high enough to cause food poisoning in humans. Multiplication and histamine formation by Proteus morganii at different temperatures show (Fig. 1 and 2) that the concentration of histamine increases rapidly once the formation has started and the level of 1000 mg/kg considered as critical may be exceeded within a short period of time. These results confirm our observations obtained during epidemiological studies of food poisoning cases that improper handling and storage of tuna meat from cans at food serving establishments may bear the risk of causing scombroid poisoning. It should be emphasized that certain cultures of Proteus morganii are able to form histamine at levels of more than 1000 mg/kg even at a temperature of 7 degree C although prolonged incubation is necessary (Fig. 1). This fact should be taken into consideration in the evaluation of results when a prolonged period of time has elapsed between sampling or consumption of the suspect food and its examination.

Animals↗

Urinary tract infection with proteus species in a teaching hospital.

This study was undertaken to assess the prevalence of urinary tract infection due to proteus species, the predisposing factors, age and sex distribution, complications and the extent of drug resistance. A total of 580 patients were studied. Proteus species comprised 33 (9%) of the total number of bacterial isolates causing urinary tract infection during a one year study period between June 1991-May 1992. Proteus urinary tract infection was found to be the most common from the age of 55 years upwards in males, while in females, they were more evenly distributed. All the proteus isolates were completely resistant to ampicillin, tetracycline and cotrimoxazole. Ofloxacin was the most potent antibiotic (94.4%) sensitive, followed by gentamicin (83.3%) sensitive. Majority of the cases occurred in surgical department with more cases being nosocomially acquired than community. All male patients had at least one predisposing factor with catheterization being the commonest. To help reduce morbidity and mortality associated with urinary tract infection due to proteus species, strict aseptic catheter techniques and more efficient infection control measures must be reinforced.

Adult↗

Studies of antibiotic resistance of rough and smooth Proteus mirabilis strains and influence of polymyxin E on their lipopolysaccharide composition.

The influence of type of bacterial culture media on antibiotic resistance of Proteus mirabilis R and S forms, was tested. P. mirabilis S1959 (S form), R45 and R110 strains (Re and Ra mutant, respectively) cultivated in media supplemented with 10% heat inactivated bovine serum were resistant to ampicillin, amoxicillin, nalidixic acid and nitroxoline. Proteus strains cultivated in media without serum were sensitive to these antibacterial agents. The presence of serum did not change the polymyxin E (colistin) resistance of there Proteus strains tested. The effects of the presence of colistin (1000 U/ml) in culture media on Proteus lipopolysaccharide composition was studied. The content of uronic acids and phosphate residues in lipopolysaccharides isolated from bacteria cultivated in the presence of colistin (LPS-col), were lower than in control LPSs. The contents of 4-amino-4-deoxy-L-arabinose decreases in S1959 LPS-col, increases in R110 LPS-col and remains unchanged in R45 LPS-col. These results indicate that the presence of colistin in cultivation media exerts an influence on the contents of charged components of LPSs isolated from polymyxin E-resistant Proteus R and S strains.

Amino Sugars↗

Investigation of the types and characteristics of the proteolytic enzymes formed by diverse strains of Proteus species.

Many diverse clinical isolates of Proteus mirabilis (48 strains), P. penneri (25), P. vulgaris biogroup 2 (48) and P. vulgaris biogroup 3 (21) from man were examined for their ability to produce proteolytic enzymes and the nature and characteristics of the proteases were studied. All the P. penneri isolates, most (94-90%) of the P. mirabilis and P. vulgaris biogroup 2 isolates, but only 71% of the P. vulgaris biogroup 3 isolates, secreted proteolytic enzymes. These were detected most readily at pH 8 with gelatin as substrate. A strong correlation was found between the ability of a strain to form swarming growth and its ability to secrete proteases. Non-swarming isolates invariably appeared to be non-proteolytic. However, some isolates, particularly of P. vulgaris biogroup 3, were non-proteolytic even when they formed swarming growth. Analysis of the secreted enzymes of the different Proteus spp. on polyacrylamide-gelatin gels under various constraints of pH and other factors showed that they were all EDTA-sensitive metalloproteinases. Analysis of the kinetics of production of the proteases revealed the formation of an additional protease of undefined type and function that was cell-associated and formed before the others were secreted. The secreted protease was subsequently modified to two isoforms whose mass (53-46 kDa) varied with the Proteus spp. and the strain. There was no evidence that the secreted proteases of strains of Proteus spp. were of types other than metalloproteinases.

Animals↗

Characterisation of the Rac/PAK pathway in Amoeba proteus.

Molecular mechanisms underlying the unique locomotion of the highly motile Amoeba proteus still remain poorly understood. Recently, we have shown that blocking the endogenous amoebal Rac-like protein(s) leads to distinct and irreversible changes in the appearance of these large migrating cells as well as to a significant inhibition of their locomotion. To elucidate the mechanism of the Rac pathway in Amoeba proteus, we tested the effects of blocking the endogenous myosin I heavy chain kinase (MIHCK), one of the Rac effectors in Acanthamoeba castellanii and Dictyostelium discoideum, with anti-MIHCK antibodies in migrating amoebae, as well as the effect of inhibiting Rac and MIHCK on the actin polymerisation process. Antibodies against A. castellanii MIHCK detected an A. proteus protein with a molecular mass (ca. 95 kDa) similar to the A. castellanii kinase. The cellular distribution of MIHCK in A. proteus was very similar to those of Rac-like protein in amoebae and MIHCK in A. castellanii. Amoebae microinjected with anti-MIHCK antibodies moved slower and protruded fewer wide pseudopodia (5-6) than the control cells (9-10), resembling to some extent the phenotype of cells microinjected with anti-Rac antibodies. The in vitro studies indicate that the A. proteus Rac-like protein, but not the MIHCK isoform, is engaged in the regulation of the nucleation step of the actin polymerisation process. These observations suggest that MIHCK may be one of the effectors for Rac in these extremely large cells.

Actins↗

ProTeus: identifying signatures in protein termini.

ProTeus (PROtein TErminUS) is a web-based tool for the identification of short linear signatures in protein termini. It is based on a position-based search method for revealing short signatures in termini of all proteins. The initial step in ProTeus development was to collect all signature groups (SIGs) based on their relative positions at the termini. The initial set of SIGs went through a sequential process of inspection and removal of SIGs, which did not meet the attributed statistical thresholds. The SIGs that were found significant represent protein sets with minimal or no overall sequence similarity besides the similarity found at the termini. These SIGs were archived and are presented at ProTeus. The SIGs are sorted by their strong correspondence to functional annotation from external databases such as GO. ProTeus provides rich search and visualization tools for evaluating the quality of different SIGs. A search option allows the identification of terminal signatures in new sequences. ProTeus (ver 1.2) is available at http://www.proteus.cs.huji.ac.il.

Databases, Protein↗

Functional morphology of the inner ear and underwater audiograms of Proteus anguinus (Amphibia, Urodela).

Octavolateral sensory organs (auditory and lateral line organs) of cave salamander Proteus anguinus are highly differentiated. In the saccular macula of the inner ear the complex pattern of hair cell orientation and the large otoconial mass enable particle displacement direction detection. Additionally, the same organ, through air cavities within the body, enables detection of underwater sound pressure changes thus acting as a hearing organ. The cavities in the lungs and mouth of Proteus are a resonators that transmit underwater sound pressure to the inner ear. Behaviourally determined audiograms indicate hearing sensitivity of 60 dB (rel. 1 microPa) at frequencies between 1 and 10 kHz. The hearing frequency range was between 10 Hz and 10 kHz. The hearing sensitivities of depigmented Proteus and black Proteus were compared. The highest sensitivities of the depigmented animals (N=4) were at frequencies 1.3-1.7 kHz and it was 2 kHz in black animals (N=1). Excellent underwater hearing abilities of Proteus are sensory adaptations to cave habitat.

Acoustic Stimulation↗

Proteus dysbioses in patients with ulcerative colitis.

In patients with ulcerative colitis Proteus could be isolated from all the gastrointestinal portions studied: mouth cavity, stomach, small intestine, small intestinal mucous membrane, and mostly from faeces. In some patients with ulcerative colitis a decrease in membraneous hydrolysis of polysaccharides was noted. Severe Proteus dysbiosis was associated with a sharp decrease in absorption processes at the small intestinal mucous membrane. The Proteus strains obtained from the patients belonged mostly to 3 serogroups: OA, OB, OC. Studies in virulent properties of Proteus in experiments with white mice gave positive results in 96% cases. Enterotoxin could be found in no case. As the result of vaccinal therapy in most patients disappearance or decrease of signs of malabsorption, increase in weight, stool normalization, and better hematological indices was manifested.

Colitis, Ulcerative↗

[Black nails caused by Proteus mirabilis].

Black nails caused by Proteus mirabilis were seen in a motor mechanic and in a petrol pump attendant. Proteus mirabilis is a gram-negative bacillus that generates hydrogen sulfide. This compound reacts with traces of metals in the nail plate such as zinc, nickel, cobalt, iron, manganese, tin, copper and lead. Metal sulfides blacken the nail plate. The protracted course of the discoloration over months corresponds to the slow reactions of metals with hydrogen sulfide. The disappearance of the blackening after topical treatment with chinosol, tincture of iodine and chloramphenicol solution supports the etiologic connection between black nails and Proteus mirabilis. Wet and dirty work encourages the colonisation of Proteus mirabilis between nail fold and nail plate.

Administration, Topical↗

Anti-Proteus antibodies in rheumatoid arthritis same-sexed sibships.

One hundred and forty-two RA patients and 121 of their healthy same-sexed siblings were tested for antibodies (Abs) to Proteus mirabilis and Escherichia coli by indirect immunofluorescence. Eighty-five individuals had active RA (CRP greater than 10 mg/l) and 57 inactive RA. The anti-Proteus Ab titre in the active RA group was significantly higher than in the inactive RA and non-RA group (P less than 0.0001 in both cases). Anti-Proteus Abs were significantly elevated in 30 individuals with active RA compared with their healthy HLA-identical same-sexed siblings (P less than 0.001). In 42 HLA-DR4 positive and 15 HLA-DR4 negative active RA patients, the Ab titre was non-significantly higher in the former group. Anti-E. coli Abs were not significantly elevated in any of the groups. Longitudinal data on 36 RA patients demonstrated a significant positive correlation between changes in CRP and changes in anti-Proteus antibody titres (r = 0.52, P less than 0.001). These observations require an explanation.

Antibodies, Bacterial↗

Cytotoxic activity of the Proteus hemolysin HpmA.

We previously showed that hpmA is the hemolysin determinant most commonly found among Proteus isolates. To assess the potential contribution of HpmA to virulence, we first characterized the toxic activities of this hemolysin. Hemolytic activity was present in total cell cultures and cell-free supernatants of Proteus clinical isolates as well as Escherichia coli containing cloned hpm genes. HpmA also possesses cytotoxic activity which was detected by a chromium release assay against a variety of target cell lines (Daudi, Raji, T24, U937, and Vero). Analysis of the dose response of bacterial cells against both T24 cells and erythrocytes showed that E. coli containing cloned hpm genes was 30-fold more cytotoxic than Proteus mirabilis BA6163. Also, 10(5)-fold more bacterial cells were needed to lyse T24 cells than to lyse erythrocytes. HpmA- mutants of two Proteus strains in which the central portion of hpmA was deleted were constructed. These HpmA- mutants, which have lost the hemolytic and cytotoxic activities exhibited by their respective parent strains, demonstrate that HpmA is needed for both of these activities. In an ascending model of murine urinary tract infection, the hpmA mutant strain WPM111 behaved no differently from its parent strain, BA6163, with respect to either the level of kidney colonization or histopathological changes in the kidney. However, WPM111 had a sixfold higher 50% lethal dose than BA6163 when injected intravenously into C3H mice.

Animals↗

Assessment of technique for rapid detection of Escherichia coli and Proteus species in urine by head-space gas-liquid chromatography.

A test depending on the production of ethanol by Escherichia coli from lactose and dimethyl disulfide by Proteus spp. from methionine in the early exponential phase of growth and the detection of these products by head-space gas-liquid chromatography has been applied to 75 specimens of urine selected to provide the most stringent trial of the test. The test was found to be rapid and reliable for the commonest findings in the microbiological examination of urine. In 3 to 4 h it detected "significant" numbers (greater than 10(5)/ml) of E. coli or of Proteus mirabilis or P. inconstans A, identified as Proteus spp., in 23 urines. It recorded the absence of infection from 32 urines containing borderline or "not significant" numbers of any organism. Significant numbers of other organisms in 13 urines were not mistaken for E. coli or Proteus spp. However, the test was less successful for some less common findings. Klebsiella ozenae in significant numbers in one urine was mistaken for E. coli. P. morganii in significant numbers in one urine was not detected. E. coli or P. mirabilis mixed with significant numbers of another organism were not detected in four out of five urines. The technique is simple and could be automated. It appears to merit more extensive trial in a hospital laboratory and further development to detect and correctly identify more species that cause urinary tract infections.

Chromatography, Gas↗

Typing of proteus from patients with bacteriuria.

Forty-five strains of Proteus mirabilis isolated from hospital in-patients, out-patients and community patients with significant bacteriuria, were investigated to determine whether a predominant type was present using the Dienes test. Antimicrobial susceptibility patterns of the 45 strains of Proteus mirabilis were also determined. There were 38 different Dienes types which shows that there is no predominant type of Proteus amongst the isolates examined in this study. This suggests that there is no predominant type of Proteus mirabilis in the community and hospital acquired bacteriuria. The antimicrobial susceptibility pattern of an isolate was not a useful marker for typing because susceptibility patterns between different Dienes types were so similar.

Adolescent↗

Serological classification and epitope specificity of Proteus penneri S29 lipopolysaccharide.

INTRODUCTION: Gram-negative bacteria of genus Proteus are common human intestinal and urinary tract pathogens. In the genus Proteus there are four clinically important named species: P. mirabilis, P. vulgaris, P. penneri, and P. hauseri, and three unnamed Proteus genomospecies: 4, 5, and 6. The clinical significance of P. penneri, described in 1982 as a new species, is poorly documented. The aim of this work is serological characterization and classification of a ceftriaxone-susceptible P. penneri S29 strain isolated from a 34-year-old patient with postneurosurgical meningitis. In this characterization we will also include a ceftriaxonresistant strain, P. penneri R15, isolated from the same patient after 12 days' treatment with ceftriaxon and other antibiotics. MATERIAL/METHODS: Rabbit polyclonal O-antisera were obtained against these two strains and purified and lipopolysaccharides (LPS) were extracted from the bacterial mass of the P. penneri S29 and R15 strains. In the serological investigations the following tests were used: enzyme immunosorbent assay (EIA), passive immunohemolysis (PIH), inhibition of these tests, absorption of rabbit O-antisera with the respective LPS, and repeated PIH, SDS/PAGE, and Western blot techniques. RESULTS: The serological studies of the LPS extracted from both P. penneri strains showed the identity of both preparations of O-polysaccharides from LPS. In P. penneri S29 O-antiserum, four different types of antibodies were described and characterized. CONCLUSIONS: Both investigated P. penneri S29 and R15 strains were classified to the Proteus O31ab serogroup.

Adult↗

[Preventive properties of the blood sera from persons vaccinated with a Proteus vaccine made from soluble antigen complexes].

In experiments of the passive protection of mice the protective properties of sera obtained from humans before and after their immunization with Proteus vaccine used as a monopreparation or in combination with staphylococcal toxoid and/or pyoimmunogen were studied. When introduced in a single subcutaneous injection, Proteus vaccine prepared from soluble antigenic complexes ensured an increase in the protective properties of sera. The second injection of the vaccine essentially enhanced the protective potency of the sera of the immunized donors. The therapeutic injection of Proteus vaccine ensured the essential increase of the protective properties of the sera. This increase could be experimentally detected within at least 25-30 days from the beginning of immunization. The immunization of volunteers with Proteus vaccine in combination with pyoimmunogen and adsorbed staphylococcal toxoid ensured the maximum increase of the protective properties of their sera.

Animals↗

[Pseudomonas aeruginosa and Proteus antigenemia and antibody formation in mono- and mixed infections in patients with suppurative inflammatory diseases].

P. aeruginosa and Proteus antigenemia and antibody production have been studied in 335 patients with purulent inflammatory diseases. The study has revealed that in the association of P. aeruginosa and Proteus with staphylococci and representatives of the family Enterobacteriaceae the level of antigenemia is considerably lower than in monoinfections or in the association of these microorganisms with streptococci. In mixed infections humoral immune response develops later than in cases of monoinfection. An important role of ecological and physiological relationships between microorganisms in the course of purulent processes and in their influence on the host has been confirmed. The use of the enzyme immunoassay in the clinical practice has made it possible to determine the etiological role of P. aeruginosa and Proteus in the development of suppurative-inflammatory diseases, to select adequate immunotherapy, including the use of anti-P. aeruginosa and anti-Proteus plasmas, and to evaluate the effectiveness of treatment.

Adolescent↗